This invention concerns a microbial process for production of compositions of matter containing isomers of jasmonic acid having the structure: 
which are flavor acceptable as well as isomers having the structure: 
which are also flavor acceptable. This invention is also concerned with the production of methyl jasmonate isomers from the aforementioned jasmonic acid isomers.
Considerable time and effort have been expended by microbiologists in the search for better processes for the production of jasmonic acid and methyl jasmonate which are flavor acceptable isomers, including those isomers defined according to the structure: 
wherein R is methyl or hydrogen. Thus, Broadbent in United Kingdom Patent Specification No. 1,286,266 published on Aug. 23, 1972 discloses and claims a process for the manufacture of jasmonic acid which comprises cultivation of the organism Lasiodiplodia theobromae in a nutrient medium containing an assimilable source of carbon and an assimilable source of nitrogen followed by isolation of the product from the culture medium. Gxc3xcnther, et al, German Democratic Republic Patent DD 279 688 published on Jun. 13, 1990 discloses a process for the production of 7-iso-jasmonic acid by strains of the organism Botryodiplodia theobromae in aerobic culture. This work is also discussed in the paper Miersch, et al, Phytochemistry, Volume 26, No. 4, pages 1037-1039, 1987, entitled xe2x80x9c(+)-7-ISO-JASMONIC ACID AND RELATED COMPOUNDS FROM BOTRYODIPLODIA THEOBROMAE.xe2x80x9d The organisms, Botryodiplodia theobromae and Lasiodermea theobromae are synonyms of Diplodia gossypina as discussed in the paper Jones, MYCOTAXON, Volume VI, No. 1 at pages 24-26, July-September 1997 (title: xe2x80x9cTHE CURRENT TAXONOMIC STATUS OF DIPLODIA GOSSYPINA.xe2x80x9d As confirmed by Hxc3xa4usler and Mxc3xcnch in the article entitled xe2x80x9cMicrobial Production of Natural Flavors,xe2x80x9d Volume 63, No. 10, ASM News at pages 551-559:
xe2x80x9cAnother plant fatty acid metabolite, jasmonic acid, an endogenous plant growth regulator with a variety of physiological functions, is produced by means of a similar metabolic pathway. After a lipoxygenase produces a hydroperoxide derivative of linolenic acid, this compound is converted to its allene oxide, which cyclizes. xcex2-Oxidation and double-bond reduction yields jasmonic acid. The methylester of jasmonic acid is not only a volatile plant hormone, possibly involved in interplant communication, but is also an important flavor and fragrance molecule that imparts a sweet-floral, jasmine-like note . . .
Otto Miersch and his collaborators at the Institute of Plant Biochemistry, Halle-Saale, Germany, who were studying fungal plant pathogens, including Botryodiplodia theobromae, discovered that such microorganisms produce jasmonic acid. The biosynthetic steps leading to jasmonic acid in this filamentous fungus are probably similar to those found in plants. Recently our laboratory, which is evaluating this strain""s capacity for producing jasmonic acid, found that B. theobromae yields only very low concentrations of jasmonic acid in liquid culture. Such findings suggest that the biosynthesis and excretion of jasmonic acid is strictly controlled during the growth cycle of this fungus on plants in its natural habitat.xe2x80x9dno bioprocesses for the high yield production of fruit flavor-acceptable jasmonic acid or methyl jasmonate exist The Hxc3xa4usler and Mxc3xcnch paper was published in October 1997. ASM News is published by the American Society for Microbiology.
Thus, in the flavor and fragance art, a need has arisen for the development and efficient high yield production of naturally occurring jasmonic acid and methyl jasmonate, which have heretofore been found to be useful and necessary for the creation of flavor formulations used in augmenting or enhancing the aroma or taste of foodstuffs, chewing gums, toothpaste, medicinal products, chewing tobaccos and smoking tobaccos and also useful in augmenting or enhancing the aroma of perfume compositions, colognes and perfumed articles (e.g., solid or liquid anionic, cationic, nonionic or zwitterionic detergents, perfumed polymers, fabric softener compositions, fabric softener articles, hair preparations, cosmetic powders and the like).
Methyl jasmonate (without indicating which isomer) is disclosed by Arctander, Perfume and Flavor Chemicals at monograph 2093 to have a xe2x80x9cPowerful floral-herbaceous, sweet-tenacious odor representing typical background notes of Jasmin absolute.xe2x80x9d Arctander further discloses that this material xe2x80x9cis an almost obvious candidate for work on improved artificial Jasmin absolute.xe2x80x9d Acree, et al, J. Agric. Food Chem., 1985, Volume 33, pages 425-427, discloses that the isomer of methyl jasmonate having the structure: 
having an optical rotation (xcex1D20) of +58 has a strong odor, whereas the other stereoisomers of (Z)-methyl jasmonate are substantially odorless.
No specific disclosures exist in the prior art showing the use of the jasmonic acid isomers of our invention defined according to the structure: 
or defined according to the structure: 
for use as flavor adjuvants (wherein R is methyl or hydrogen). Furthermore, nothing in the prior art discloses the stereoisomer of the compound having the structure: 
particularly the stereoisomer having the optical rotation (xcex1D20) of +58xc2x0.
Our invention relates to a bioprocess for the high yield production of food flavor acceptable jasmonic acid and methyl jasmonate, a novel jasmonic acid isomer produced thereby and uses thereof. The process of our invention yields at least 5% of the xe2x80x9ccisxe2x80x9d isomer defined according to the structure: 
(wherein R is hydrogen or methyl) or the xe2x80x9ccisxe2x80x9d isomer defined according to the structure: 
(wherein R is hydrogen or methyl). Compositions containing at least 98% of the isomer having the structure: 
having an xcex1D20 of +58xc2x0 are novel. Furthermore, compounds having the structure: 
wherein R is methyl or hydrogen having an xcex1D20 of +58xc2x0 are also novel.
The process of our invention comprises the cultivation under aerobic conditions of one or more specific strains of Diplodia gossypina in a nutrient medium followed by either (1) isolation of the jasmonic acid or (2) esterification of the jasmonic acid to form methyl jasmonate followed by isolation of the methyl jasmonate. More specifically, our process comprises cultivation under aerobic conditions of a strain of Diplodia gossypina organism selected from the group consisting of:
(i) Diplodia gossypina ATCC 10936;
(ii) Diplodia gossypina ATCC 20575;
(iii) Diplodia gossypina NRRL 25011; and
(iv) Diplodia gossypina ATCC 36037
in a nutrient medium containing an assimilable source of carbon and an assimilable source of nitrogen followed by isolation of the jasmonic acid product from the culture medium or followed by esterification of the jasmonic acid to form methyl jasmonate and then followed by isolation of the methyl jasmonate product from the reaction mass.
In one embodiment of the process of our invention, the compound, 10-oxo-trans-8-decenoic acid having the structure: 
is intimately admixed in the nutrient medium in an amount of from about 0.7 up to about 10 ppm (parts per million) by weight of the nutrient medium prior to the cultivation step.
The 10-oxo-trans-8-decenoic acid having the structure: 
is disclosed in U.S. Pat. No. 5,681,738 issued on Oct. 28, 1997 to be a fungal growth hormone to stimulate mycelial growth of cultivated mushrooms. The specification of U.S. Pat. No. 5,681,738 issued on Oct. 28, 1997 is incorporated by reference herein. The bioprocess of our invention is shown according to the reactions: 
wherein D.G. strain is intended to mean xe2x80x9cstrain of Diplodia gossypina.xe2x80x9d
The esterification reaction following the bioprocess reaction of our invention is shown as follows: 
Thus, the assimilable carbon source for our invention has been found to be glucose. The preferred assimilable nitrogen source is sodium nitrate.
Preferably, the aeration rate range for carrying out the cultivation step of our invention is from about 0.5 v/v/m (liters air per liter fermentation mass per minute) up to about 1.0 v/v/m.
Furthermore, it is preferable that the culture medium contain magnesium ion, for example, magnesium ion in the form of magnesium sulfate heptahydrate having the formula:
MgSO4.7H2O
with the magnesium ion concentration ([Mg++]) being in the range of from about 6xc3x9710xe2x88x923 up to about 10.5xc3x9710xe2x88x923 gram moles per liter. Thus, for example, the concentration range of magnesium sulfate heptahydrate in grams per liter preferably is in the range of from about 1.5 up to about 2.5 grams per liter of magnesium sulfate heptahydrate.
The cultivation step of the process of our invention may take place under liquid surface culture conditions or may take place under submerged liquid culture conditions. The incubation time is from about 3 up to about 15 days.
The pH range during the fermentation is from about 4.5 up to about 9, with a preferred pH of about 6.
The temperature for the fermentation reaction may vary between about 20xc2x0 C. up to about 35xc2x0 C., with a preferred temperature range for the fermentation being from about 25xc2x0 C. up to about 32xc2x0 C.
We have found that the preferred strain of Diplodia gossypina is the Diplodia gossypina ATCC 10936.
Along with the desired xe2x80x9ccisxe2x80x9d jasmonic acid isomers having the structures: 
prepared according to the reactions: 
other jasmonic acid isomers defined according to the structures: 
for example, are also produced in very small quantities when using the strains of Diplodia gossypina: 
(i) Diplodia gossypina ATCC 10936;
(ii) Diplodia gossypina ATCC 20575;
(iii) Diplodia gossypina NRRL 25011; and
(iv) Diplodia gossypina ATCC 36037.
The inventive process may be conducted in a batch or continuous mode of operation. In a batch fermentation, the nutrient broth, culture and substrate, are combined and fermented until the jasmonic acid concentration becomes constant. In a continuous process, the substrate in the nutrient broth may be continuously recirculated through a fermentation reactor, with the provision that substrate and product are respectively added and removed from the recirculating broth.
In carrying out the present invention, cultivation and fermentive incubation of the Diplodia gossypina fungus are accomplished in an aqueous medium in the presence of the usual nutrient substances (preferably using sodium nitrate, magnesium sulfate heptahydrate and glucose). A suitable medium thus is one which contains carbon sources, nitrogen sources, inorganic salts and growth factors. Additional examples of inorganic salts include the phosphate salts of magnesium and the phosphate and sulfate salts of sodium, calcium and potassium. These nutrients may be supplemented with, for example, one or more vitamins of the xe2x80x9cBxe2x80x9d group and one or more trace minerals such as iron, manganese, cobalt and copper, as desired.
For the nutrient broth, it is preferred to utilize glucose (as stated, supra) at a concentration of from about 2 up to about 20 weight percent, preferably at about 10 weight percent. It is also preferred to employ xe2x80x9cBxe2x80x9d vitamins either as a separate supplement or in the form of a yeast extract. The kind and amounts of these additives can be determined by applying the general knowledge in the art for the cultivation of microorganisms.
In a typical procedure, one of the Diplodia gossypina organisms:
(i) Diplodia gossypina ATCC 10936;
(ii) Diplodia gossypina ATCC 20575;
(iii) Diplodia gossypina NRRL 25011; or
(iv) Diplodia gossypina ATCC 36037
is first cultivated in inoculum quantities to produce a mature culture in nutrient broth. The culture is inoculated into a fermentor nutrient broth and allowed to establish itself The substrate is then added and fermentation is continued until a steady concentration of jasmonic acid is present.
The cultivation and fermentative incubation of the Diplodia gossypina fungus can be carried out as a stationary culture or as a submerged culture (e.g., shake-flask fermentor) under aerobic conditions. Cultivation and incubation may proceed as stated, supra, in a pH range of from about 4.5 up to about 9, preferably at 6. The pH may be regulated by the addition of an inorganic or organic acid or base such as hydrochloric acid, acetic acid, sodium hydroxide, calcium carbonate, ammonia, ion-exchange resins or by the addition of a buffer such as phosphate or phthalate. The incubation temperature as stated, supra, is suitably maintained between about 20xc2x0 C. up to about 35xc2x0 C., with a range of from about 25xc2x0 C. up to about 32xc2x0 C. being preferred.
In accordance with another typical procedure of the present invention, the process is conveniently carried out by adding the substrate to the culture medium at the onset of cultivation under aerobic conditions. Alternatively, the substrate may be added either alone or in combination with another carbon source, such as xylose during fermentative incubation or when cultivation is complete. It is preferable to add the substrate to the culture medium during the period of from 4 up to 24 hours after the growth of the culture in the fermentative broth has commenced. Desirable results can be obtained when the substrate is added continuously over the entire fermentation after an initial fungal (Diplodia gossypina) cultivation period of from 3 up to 12 hours. A preferred feed rate for this continuous addition is from about 0.01 up to 1 gram per hour per liter, with a preferred range of from 0.6 up to 0.8 grams per hour per liter. The concentration of the substrate in the medium may vary depending on the conditions employed. In practice, the concentration of the substrate in the medium may conveniently vary from 0.01% up to about 10%, with a preferable concentration of about 1% by weight consistent with the manner in which it is added to the culture.
The present invention provides unexpectedly high yields of jasmonic acid, e.g., from about 0.8 up to about 1.5 grams per liter.
Another aspect of our invention is the production of methyl jasmonate isomers having the structure: 
and having the structure: 
In this case, the isolation step comprises the steps of:
(a) extraction of the jasmonic acid from the fermentation broth with an extraction solvent such as ethyl acetate to form a jasmonic acid extract;
(b) concentration of the jasmonic acid extract whereby the extraction solvent is stripped;
(c) esterification of the resulting jasmonic acid with methyl alcohol whereby the methyl jasmonate is formed according to one of the reactions: 
(d) concentration of the resulting methyl jasmonate.
Additional steps of fractionation of the resulting concentrate with, for example, silica gel, in order to effect isomer separation and then collecting the 98% xe2x80x9ccisxe2x80x9d isomer are preferred.
The jasmonic acid derivative(s) defined according to the structure: 
or according to the structure: 
(wherein R is methyl or hydrogen) and one or more auxiliary perfume ingredients including, for example, hydrocarbons, alcohols, ketones (other than the jasmonic acid derivatives of our invention), aldehydes, nitrites, esters (other than the jasmonic acid derivatives of our invention), ethers, synthetic essential oils and natural essential oils, may be admixed so that the combined odors of the individual components produce a pleasant and desired fragrance, particularly and preferably in the floral area (e.g., jasmine and jasmine/rose aromas). Such perfume compositions usually contain (a) the main note or the xe2x80x9cbouquetxe2x80x9d or foundation stone of the composition; (b) modifiers which round off and accompany the main note; (c) fixatives which include odorous substances which lend a particular note to the perfume throughout all stages of evaporation, and substances which retard evaporation; and (d) topnotes which are usually low-boiling, fresh-smelling materials.
In perfume compositions, it is the individual components which contribute to their particular olfactory characteristics; however, the overall sensory effect of the perfume composition will be at least the sum total of the effects of each of the ingredients. Thus, one or more of the jasmonic acid derivative(s) of our invention can be used to alter, modify or enhance the aroma characteristics of a perfume composition, for example, by utilizing or moderating the olfactory reaction contributed by another ingredient in the composition. Thus, a suitable mixture would be that of the cis isomers of jasmonic acid and methyl jasmonate, to wit: the mixture of the compounds having the structures: 
or the mixture of compounds having the structures: 
The amount of jasmonic acid derivative(s) of our invention, which will be effective in perfume compositions as well as in perfumed articles and colognes, depends upon many factors including the other ingredients, their amounts and the side effects which are desired. It has been found that perfume compositions containing as little as 0.005% of the jasmonic acid derivative(s) or even less (e.g., 0.002%) can be used to impart powerful long lasting jasmine, floral-herbaceous aromas with sweet-herbaceous, green-woody topnotes to soaps, cosmetics, detergents including anionic, cationic, nonionic and zwitterionic solid or liquid detergents, perfumed polymers and other products. The amounts employed can range up to 70% of the fragrance components and will depend upon considerations of cost, nature of the end product, the effect desired on the finished product and the particular fragrance sought.
The jasmonic acid derivative(s) of our invention are useful (taken alone or taken together with other ingredients in perfume compositions) in detergents, soaps, space odorants and deodorants, perfumes, colognes, toilet waters, bath preparations, hair preparations such as lacquers, brilliantines, pomades and shampoos; cosmetic preparations such as creams, deodorants, hand lotions and sun screens; powders such as talcs, dusting powders, face powders and the like.
As little as 0.25% of the jasmonic acid derivative(s) will suffice to impart an intense and long lasting jasmine, floral-herbaceous aroma with sweet-herbaceous, green-woody topnotes to floral perfume formulations. Generally no more than 5% of the jasmonic acid derivative(s) based on the ultimate end product are required to be used in the perfume compositions.
Furthermore, as little as 0.25% of the jasmonic acid derivative(s) will suffice to impart such aromas to perfumed articles per se, whether in the presence of other perfume materials or whether used by themselves. Thus, the range of use of the jasmonic acid derivative(s) of our invention in perfumed articles, e.g., perfumed polymers and solid or liquid anionic, cationic, nonionic or zwitterionic solid or liquid detergents, may vary from 0.25% up to about 5% by weight based on the total weight of the perfumed article.
In addition, the perfume composition or fragrance composition of our invention can contain a vehicle or carrier for the jasmonic acid derivative(s). The vehicle can be a liquid such as a nontoxic alcohol, e.g., ethanol; a nontoxic glycol, e.g., propylene glycol; or the like. The carrier can also be an absorbent solid such as a gum (e.g., gum arabic, xanthan gum or guar gum) or components for encapsulating the composition by means of coacervation (such as gelatin) or by means of formation of a polymer around a liquid center (as by using a urea formaldehyde prepolymer to form a polymeric capsule around a perfume composition center).
It will be appreciated from the present disclosure that the jasmonic acid derivative(s) according to the present invention can be used to alter, vary, fortify, modify, enhance or otherwise improve the flavor of a wide variety of materials which are ingested, consumed or otherwise organoleptically sensed.
The terms xe2x80x9calterxe2x80x9d and xe2x80x9cmodifyxe2x80x9d in their various forms will be understood herein to mean the supplying or imparting of a flavor character or note to an otherwise bland, relatively tasteless substance, or augmenting an existing flavor characteristic where the natural flavor is deficient in some regard or supplementing the existing flavor impression to modify its organoletpic character.
The term xe2x80x9cenhancexe2x80x9d is intended herein to mean the intensification (by use of the jasmonic acid derivative of our invention) of a flavor or aroma note or nuance in a tobacco flavor or foodstuff or perfume composition or perfumed article without changing the quality of said note or nuance.
A xe2x80x9cflavoring compositionxe2x80x9d is taken to mean one which contributes a part of the overall flavor impression by supplementing or fortifying a natural or artificial flavor in a material, or one which supplies substantially all the flavor and/or aroma character to a consumable article.
The term xe2x80x9cfoodstuffxe2x80x9d as used herein includes both solid and liquid ingestible materials for man or animals, which materials usually do, but need not, have nutritional value. Thus, foodstuffs include meats, gravies, soups, convenience foods, malt, alcoholic and other beverages, milk and dairy products, seafoods, including fish, crustaceans, mollusks and the like, candies, vegetables, cereals, soft drinks, snacks, dog and cat foods, other veterinary products and the like. The jasmonic acid derivative(s) of our invention are also useful in tobacco flavorants and tobacco enhancers.
The term xe2x80x9ctobaccoxe2x80x9d will be understood herein to mean natural products such as, for example, Burley, Turkish tobacco, Maryland tobacco, flue-cured tobacco and the like, including tobacco-like or tobacco-based products such as reconstituted or homogenized leaf and the like, as well as tobacco substitutes intended to replace natural tobacco such as lettuce and cabbage leaves and the like. The tobaccos and tobacco products in which the jasmonic acid derivative(s) of our invention are useful include those designed or used for smoking such as in cigarettes, cigar and pipe tobacco, as well as products such as snuff, chewing tobacco and the like.
When the jasmonic acid derivative(s) of this invention are used in a flavoring composition, they can be combined with conventional flavoring materials or adjuvants. Such co-ingredients or flavor adjuvants are well known in the art for such use and have been extensively described in the literature. Requirements of such adjuvant materials are: (1) that they be non-reactive with the jasmonic acid derivative(s) of our invention; (2) that they be organoleptically compatible with the jasmonic acid derivative(s) of our invention whereby the flavor of the ultimate consumable material to which the jasmonic acid derivative(s) are added is not detrimentally affected by the use of the adjuvant; and (3) that they be ingestibly acceptable and thus nontoxic or otherwise non-deleterious. Apart from these requirements, conventional materials can be used and broadly include other flavor materials, vehicles, stabilizers, thickeners, surface active agents, conditioners and flavor intensifiers.
Such conventional flavoring materials include saturated fatty acids, unsaturated fatty acids and amino acids; alcohols including primary and secondary alcohols, esters, carbonyl compounds including ketones (other than the jasmonic acid derivatives of our invention) and aldehydes; lactones; other cyclic organic materials including benzene derivatives, alicyclic compounds, heterocyclics such as furans, pyridines, pyrazines and the like; sulfur-containing compounds including thiols, sulfides, disulfides and the like; proteins; lipids, carbohydrates; so-called flavor potentiators such as monosodium glutamate; magnesium glutamate, calcium glutamate, guanylates and inosinates; natural flavoring materials such as cocoa, vanilla and caramel; essential oils and extracts such as anise oil, clove oil and the like and artificial flavoring materials such as vanillin, ethyl vanillin and the like.
Specific preferred flavor adjuvants are as follows:
anise oil;
ethyl-2-methyl butyrate;
vanillin;
cis-3-heptenol;
cis-3-hexenol;
trans-2-heptenal;
butyl valerate;
2,3-diethyl pyrazine;
methyl cyclopentenolone;
benzaldehyde;
valerian oil;
3,4-dimethoxyphenol;
amyl acetate;
amyl cinnamate;
xcex3-butyryl lactone;
furfural;
trimethyl pyrazine;
phenyl acetic acid;
isovaleraldehyde;
ethyl maltol;
ethyl vanilin;
ethyl valerate;
ethyl butyrate;
cocoa extract;
coffee extract;
peppermint oil;
spearmint oil;
clove oil;
anethol;
cardamom oil;
wintergreen oil;
cinnamic aldehyde;
ethyl-2-methyl valerate;
xcex3-hexenyl lactone;
2,4-decadienal;
2,4-heptadienal;
methyl thiazole alcohol (4-methyl-5-xcex2-hydroxyehtyl thiazole);
2-methyl butanethiol;
4-mercapto-2-butanone;
3-mercapto-2-pentanone;
1-mercapto-2-propane;
benzaldehyde;
furfural;
furfuryl alcohol;
2-mercapto propionic acid;
alkyl pyrazine;
methyl pyrazine;
2-ethyl-3-methyl pyrazine;
tetramethyl pyrazine;
polysulfides;
dipropyl disulfide;
methyl benzyl disulfide;
alkyl thiophene;
2,3-dimethyl thiophene;
5-methyl furfural;
acetyl furan;
2,4-decadienal;
guiacol;
phenyl acetaldehyde;
xcex2-decalactone;
d-limonene;
acetoin;
amyl acetate;
maltol;
ethyl butyrate;
levulinic acid;
piperonal;
ethyl acetate;
n-octanal;
n-pentanal;
n-hexanal;
diacetyl;
monosodium gulatamate;
monopotassium glutamate;
sulfur-containing amino acids, e.g., cysteine;
hydrolyzed vegetable protein;
2-methylfuran-3-thiol;
2-methyldihydrofuran-3-thiol;
2,5-dimethylfuran-3-thiol;
hydrolyzed fish protein;
tetramethyl pyrazine;
propylpropenyl disulfide;
propylpropenyl trisulfide;
diallyl disulfide;
diallyl trisulfide;
dipropenyl disulfide;
dipropenyl trisulfide;
4-methyl-2-[(methylthio)-ethyl]-1,3-dithiolane;
4,5-dimethyl-2-(methylthiomethyl)-1,3-dithiolne; and
4-methyl-2-(methylthiomethyl)- 1,3-dithiolane.
The jasmonic acid derivative(s) of our invention or compositions incorporating them, as mentioned above, can be combined with one or more vehicles or carriers for adding them to the particular product. Vehicles can be edible or otherwise suitable materials such as ethyl alcohol, propylene glycol, water and the like, as described supra. Carriers include materials such as gum arabic, carrageenan, xanthan gum, guar gum and the like.
The jasmonic acid derivative(s) prepared according to our invention can be incorporated with the carriers by conventional means such as spray-drying, drum-drying and the like. Such carriers can also include materials for coacervating the jasmonic acid derivative(s) of our invention to provide encapsulated products, as set forth supra. When the carrier is an emulsion, the flavoring composition can also contain emulsifiers such as mono- and diglycerides or fatty acids and the like. With these carriers or vehicles, the desired physical form of the compositions can be prepared.
The quantity of jasmonic acid derivative(s) utilized should be sufficient to impart the desired flavor characteristic to the product, but on the other hand, the use of an excessive amount of the jasmonic acid derivative(s) is not only wasteful and uneconomical, but in some instances, too large a quantity may unbalance the flavor or other organoleptic properties of the product consumed. The quantity used will vary depending upon the ultimate foodstuff; the amount and type of flavor initially present in the foodstuff; the further process or treatment steps to which the foodstuff will be subjected; regional and other preference factors; the type of storage, if any, to which the product will be subjected; and the preconsumption treatment such as baking, frying and so on, given to the product by the ultimate consumer. Accordingly, the terminology xe2x80x9ceffective amountxe2x80x9d and xe2x80x9csufficient amountxe2x80x9d is understood in the context of the present invention to be quantitatively adequate to alter the flavor of the foodstuff.
It is accordingly preferred that the ultimate composition contain from about 0.1 parts per million (ppm) up to about 500 ppm of the jasmonic acid derivative(s).
The jasmonic acid derivative(s) of our invention when utilized in flavoring compositions can be varied over a wide range depending upon the particular flavor nuances desired to be added to the foodstuff. Thus, the amounts of jasmonic acid derivative(s) of our invention may be contained in flavoring materials from about 1 ppm up to about 50% by weight of the flavoring composition. Indeed, the compounds having the structures: 
may be utilized in raspberry flavors (e.g., for use in raspberry-flavored yogurt, for example) at levels of between about 1% and about 50%. Such materials also have utility in flavorings for salad dressings, particularly in such cuisines as xe2x80x9cThaixe2x80x9d cuisine.
According to another aspect of our invention, an organoleptically improved smoking tobacco product and additives therefor as well as methods of making the same which overcome specific problems heretofore encountered in which specific Turkish, oriental-like aromas prior to smoking and improved Turkish, oriental aromas on smoking in the mainstream and the side stream are created or enhanced or modified or augmented and may be readily controlled and maintained at the desired uniform level regardless of variations in the tobacco components of the blend. In particular, low grade Virginia-type tobaccos may be upgraded using the jasmonic acid derivative(s) of our invention.
This invention further provides improved tobacco additives and methods whereby various desirable natural aromatic Turkish tobacco flavoring characteristics with oriental notes may be imparted to smoking tobacco products and may be readily varied and controlled to produce the desired uniform flavoring characteristics.
In carrying out this aspect of our invention, we add to smoking tobacco materials or a suitable substitute therefor (e.g., dried lettuce leaves) an aroma and flavor additive containing as an active ingredient one or more of the jasmonic acid derivative(s) of our invention.
In addition to the jasmonic acid derivative(s) of our invention, other flavoring and aroma additives may be added to the smoking tobacco material or substitute therefor, either separately or in admixture with the jasmonic acid derivative(s) of our invention as follows:
xcex2-Ethyl-cinnamaldehyde;
Eugenol;
Dipentene;
xcex2-Damascenone;
Maltol;
Ethyl maltol;
Delta-Undecalactone;
Delta-Decalactone;
Benzaldehyde;
Amyl acetate;
Ethyl butyrate;
Ethyl valerate;
Ethyl acetate;
2-Hexenol-1;
2-Methyl-5-isopropyl-1,3-nonadiene-8-one;
2,6Dimethyl-1,6-undecadiene-10-one;
2-ethyl-5-isopropyl acetophenone;
2-Hydroxy-2,5,5,8a-tetramethyl-1-(2-hydroxyethyl)-decahydrononaphtalene;
Dodcahydro-3a,6,6,9a-tetramethyl naphthol(2,1-b) furan;
4-Hydroxy hexanoic acid, xcex3-lactone; and
Polyisoprenoid hydrocarbons defined in Example V of U.S. Pat. No. 3,589,372 issued on Jun. 29, 1971.
Celery seed oil;
Coffee extract;
Bergamot oil;
Cocoa extract;
Nutmeg oil; and
Origanum oil.
An aroma and flavoring concentrate containing one or more of the jasmonic acid derivative(s) of our invention and, if desired, one or more of the above-indicated additional flavoring additives may be added to the smoking tobacco material, to the filter or to the leaf or paper wrapper. The smoking tobacco material may be shredded, cured, cased and blended tobacco material or reconstituted tobacco material or tobacco substitutes (e.g., lettuce leaves) or mixtures thereof. The proportions of flavoring additives may be varied in accordance with taste, but insofar as enhancement or the imparting of oriental and/or Turkish tobacco notes, we have found that satisfactory results are obtained if the proportion by weight of the sum total of jasmonic acid derivative(s) to smoking tobacco material is between 50 ppm and 1,500 ppm (0.005%-0.15%) of the active ingredients to the smoking tobacco material. We have further found that satisfactory results are obtained if the proportion by weight of the sum total of jasmonic acid derivative(s) used to flavoring material is between 500 and 15,000 ppm (0.05%-1.5%).
Any convenient method for incorporating the jasmonic acid derivative(s) into the tobacco product may be employed. Thus, the jasmonic acid derivative(s) taken alone or along with other flavoring additives may be dissolved in a suitable solvent such as ethanol, diethyl ether and/or other organic solvents, and the resulting solution may either be spread onto the cured, cased and blended tobacco material or the tobacco material may be dipped into such solution. Under certain circumstances, a solution of the jasmonic acid derivative(s) taken alone or taken further together with flavoring additives as set forth above may be applied by means of a suitable applicator such as a brush or roller on the paper or leaf wrapper for the smoking product, or it may be applied to the filter by either spraying or dipping or coating.
Furthermore, it will be apparent that only a portion of the tobacco or substitute therefor need be treated, and the thus-treated tobacco may be blended with other tobaccos before the ultimate tobacco product is formed. In such cases, the tobacco treated may have the jasmonic acid derivative(s) in excess of the amounts or concentrations above indicated so that when blended with other tobaccos, the final product will have the percentage within the indicated range.
In accordance with one specific example of our invention, an aged, cured and shredded domestic Virginia tobacco is sprayed with a 20% alcohol solution of the compound having the structure: 
optical rotation (xcex1D20) equal +58xc2x0 on a dry basis. Thereafter, the alcohol is removed by evaporation and the tobacco is manufactured into cigarettes by the usual techniques. The cigarette, when treated as indicated, has a desired and pleasing aroma which is detectable in the mainstream and the side stream when the cigarette is smoked. The aroma is described as being sweeter with pronounced Turkish/oriental characteristics and with improved body and enhanced tobacco character with subsidiary xe2x80x9cfloral,xe2x80x9d xe2x80x9cjasmonicxe2x80x9d nuances in the mainstream and the side stream. In addition, interesting amber nuances are imparted.
While our invention is particularly useful in the manufacture of smoking tobacco such as cigarette tobacco, cigar tobacco and pipe tobacco, other tobacco products formed from sheeted tobacco dust or fines may also be used. Likewise, the jasmonic acid derivative(s) of our invention can be incorporated with materials such as filter tip materials, seam paste, packaging materials and the like, which are used along with tobacco to form a product adapted for smoking. Furthermore, the jasmonic acid derivative(s) can be added to certain tobacco substitutes of natural or synthetic origin (e.g., dried lettuce leaves) and, accordingly, by the term xe2x80x9ctobaccoxe2x80x9d as used throughout this specification, is meant any composition intended for human consumption by smoking or otherwise when composed of tobacco plant parts or substitute material or both. Thus, chewing tobacco is also included in the foregoing meaning of xe2x80x9ctobacco.xe2x80x9d
During the fermentation reaction of our invention, two byproducts are also formed, to wit, the compound having the structure: 
and the compound having the structure: 
the compound having the structure: 
formed via the reaction: 
It will be understood that each of the compounds having the structures: 
may be retained with the jasmonic acid derivative(s) of our composition for their flavor or fragrance utilities. Thus, in addition to the additives as set forth, supra, the compounds having the structures: 
may be xe2x80x9cretainedxe2x80x9d and not xe2x80x9cseparatedxe2x80x9d in the isolation of the jasmonic acid derivative(s) prior to the jasmonic acid derivative(s) being utilized in flavor or fragrance formulations.
More specifically, our invention also contemplates mixtures of derivatives having the structures: 
(wherein R is methyl or hydrogen) taken further together with the compounds having the structures: 